U.S. patent application number 15/527518 was filed with the patent office on 2017-11-09 for agent for preventing or improving symptoms caused by imbalance of sex hormones.
The applicant listed for this patent is MORINAGA MILK INDUSTRY CO., LTD.. Invention is credited to Eriko MISAWA, Marie SAITO, Miyuki TANAKA, Ruiqing YAO.
Application Number | 20170319599 15/527518 |
Document ID | / |
Family ID | 56074501 |
Filed Date | 2017-11-09 |
United States Patent
Application |
20170319599 |
Kind Code |
A1 |
MISAWA; Eriko ; et
al. |
November 9, 2017 |
AGENT FOR PREVENTING OR IMPROVING SYMPTOMS CAUSED BY IMBALANCE OF
SEX HORMONES
Abstract
Provided is a means with which it is possible to efficiently
prevent or improve symptoms caused by an imbalance of sex hormones
with as little pain as possible, and which can safely be taken
daily. A compound selected from the group consisting of a lophenol
compound and a cyclolanostane compound is used as an active
ingredient of an agent for preventing or improving symptoms caused
by an imbalance of sex hormones.
Inventors: |
MISAWA; Eriko; (Kanagawa,
JP) ; YAO; Ruiqing; (Kanagawa, JP) ; SAITO;
Marie; (Kanagawa, JP) ; TANAKA; Miyuki;
(Kanagawa, JP) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
MORINAGA MILK INDUSTRY CO., LTD. |
Tokyo |
|
JP |
|
|
Family ID: |
56074501 |
Appl. No.: |
15/527518 |
Filed: |
November 27, 2015 |
PCT Filed: |
November 27, 2015 |
PCT NO: |
PCT/JP2015/083475 |
371 Date: |
May 17, 2017 |
Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61P 3/00 20180101; A61P
5/30 20180101; A23L 33/30 20160801; A61P 5/24 20180101; A61K 45/06
20130101; A61P 19/00 20180101; A23V 2002/00 20130101; A61K 9/0095
20130101; A23L 2/52 20130101; A61K 31/575 20130101; A61P 25/24
20180101; A61P 25/00 20180101; A61K 9/0056 20130101; A23L 33/10
20160801 |
International
Class: |
A61K 31/575 20060101
A61K031/575; A23L 33/00 20060101 A23L033/00; A23L 33/10 20060101
A23L033/10; A23L 2/52 20060101 A23L002/52; A61K 9/00 20060101
A61K009/00; A61K 9/00 20060101 A61K009/00 |
Foreign Application Data
Date |
Code |
Application Number |
Nov 28, 2014 |
JP |
2014-242165 |
Claims
1-10. (canceled)
11. A method for preventing or improving symptoms caused by an
imbalance of sex hormones, comprising administering a compound
selected from the group consisting of a lophenol compound and a
cyclolanostane compound to a subject in need of prevention or
improvement of symptoms caused by an imbalance of sex hormones.
12. The method for preventing or improving symptoms caused by an
imbalance of sex hormones according to claim 11, comprising
administering a composition containing the compound at a total
amount of 0.00001% by mass or more.
13. The method for preventing or improving symptoms caused by an
imbalance of sex hormones according to claim 11, wherein the sex
hormones are female hormones.
14. The method for preventing or improving symptomscaused by an
imbalance of sex hormones according to claim 12, wherein the sex
hormones are female hormones.
15. The method for preventing or improving symptoms caused by an
imbalance of sex hormones according to claim 11, wherein the method
is for preventing or improving the symptoms not through the sex
hormone-like action.
16. The method for preventing or improving symptoms caused by an
imbalance of sex hormones according to claim 12, wherein the method
is for preventing or improving the symptoms not through the sex
hormone-like action.
17. The method for preventing or improving symptoms caused by an
imbalance of sex hormones according to claim 13, wherein the method
is for preventing or improving the symptoms not through the sex
hormone-like action.
18. The method for preventing or improving symptoms caused by an
imbalance of sex hormones according to claim 11, wherein the
symptom caused by an imbalance of sex hormones is bone
dysbolism.
19. The method for preventing or improving symptoms caused by an
imbalance of sex hormones according to claim 12, wherein the
symptom caused by an imbalance of sex hormones is bone
dysbolism.
20. The method for preventing or improving symptoms caused by an
imbalance of sex hormones according to claim 13, wherein the
symptom caused by an imbalance of sex hormones is bone
dysbolism.
21. The method for preventing or improving symptoms caused by an
imbalance of sex hormones according to claim 15, wherein the
symptom caused by an imbalance of sex hormones is bone
dysbolism.
22. The method for preventing or improving symptoms caused by an
imbalance of sex hormones according to claim 11, wherein the
symptom caused by an imbalance of sex hormones is a
neuropsychiatric symptom.
23. The method for preventing or improving symptoms caused by an
imbalance of sex hormones according to claim 12, wherein the
symptom caused by an imbalance of sex hormones is a
neuropsychiatric symptom.
24. The method for preventing or improving symptoms caused by an
imbalance of sex hormones according to claim 13, wherein the
symptom caused by an imbalance of sex hormones is a
neuropsychiatric symptom.
25. The method for preventing or improving symptoms caused by an
imbalance of sex hormones according to claim 15, wherein the
symptom caused by an imbalance of sex hormones is a
neuropsychiatric symptom.
26. The method for preventing or improving symptoms caused by an
imbalance of sex hormones according to claim 12, wherein the
composition is food or drink.
27. The method for preventing or improving symptoms caused by an
imbalance of sex hormones according to claim 14, wherein the
composition is food or drink.
28. The method for preventing or improving symptoms caused by an
imbalance of sex hormones according to claim 16, wherein the
composition is food or drink.
29. The method for preventing or improving symptoms caused by an
imbalance of sex hormones according to claim 19, wherein the
composition is food or drink.
30. The method for preventing or improving symptoms caused by an
imbalance of sex hormones according to claim 23, wherein the
composition is food or drink.
Description
TECHNICAL FIELD
[0001] The present invention relates to an agent for preventing or
improving symptoms due to an imbalance of sex hormones.
BACKGROUND ART
[0002] Hormones are chemical substances which are secreted from
endocrine glands, act on target sites in a body through blood, and
control activities thereof. Hormones are circulated in the whole
body, but many hormones act only on specific organs or tissues. At
target sites, a receptor undergoing stimulation of a hormone
exists, and transmits information for causing specific action by
binding of the hormone thereto. Hormones influence growth,
development, reproduction and the like, and at the same time,
participate in energy metabolism and homeostasis such as
maintenance of the concentrations of blood components, and control
the functions of organs in the whole body. Since hormones cause a
very great response at an extremely small amount, when the balance
thereof is disturbed, a variety of symptoms and diseases appear
(Non-Patent Document 1).
[0003] A female menopausal disorder results from disturbance in the
hormone balance which is caused by deterioration of the ovary
function and decrease in a secretion amount of estrogen in a term
of transfer from a reproductive stage to a non-productive stage.
Symptoms caused by the menopausal disorder include vasomotor
nervous symptoms (glow, hot flash, sweating, coldness, palpitation,
short breath, edema), neuropsychiatric symptoms (headache,
dizziness, insomnia, anxiety, irritation, depressive state,
tinnitus, deafness, lightheadedness, stress), motor organ symptoms
(lumbago, shoulder stiffness, arthralgia, myalgia, bone dysbolism),
digestive organ symptoms (anorexia, nausea, diarrhea, constipation,
thirst, dull feeling in the stomach, heartburn), systemic symptoms
(malaise), urinary symptoms (pollakiuria, residual urine,
micturition pain), reproduction organ symptoms (menstrual disorder,
reduction in sexual desire), sensory symptoms (numbness,
desensitization, hypersensitivity, low vision), and skin symptoms
(skin drying, itching, wrinkle, dullness). Further, it has been
elucidated that postmenopausal females are more likely to develop
hypertension, diabetes, obesity, osteoporosis, autonomic ataxia,
unidentified complaint, dementia and the like (Non-Patent Document
2).
[0004] On the other hand, increase in the menopausal disorder has
been also reported in males, and many of symptoms thereof are said
to be due to reduction in the testosterone concentration in blood
involved in a variety of neuropsychiatric symptoms. In a male
menopausal disorder, various symptoms such as vasomotor nervous
symptoms (glow, hot flash, sweating, coldness, palpitation),
neuropsychiatric symptoms (headache, dizziness, insomnia, anxiety,
depressive state, tinnitus, dyspnea), motor organ symptoms
(shoulder stiffness, arthralgia, myalgia), digestive organ symptoms
(anorexia, constipation, lumbago, mouth dryness), systemic symptoms
(malaise), urinary symptoms (pollakiuria, residual urine, reduction
in urinary stream), reproduction organ symptoms (reduction in
sexual desire), sensory symptoms (numbness, desensitization), and
the like are known (Non-Patent Document 3), like females. Further,
there is a report that also in males, reduction in estrogen
together with reduction in testosterone occur, thus, osteoporosis
is developed (Non-Patent Document 4).
[0005] Like this, in both males and females, disturbance in the sex
hormone balance is generated due to influence of deterioration in
the function of organs due to aging in a menopausal stage, and also
in females in young generation, an imbalance of sex hormones is
generated in some cases. As symptom caused by such an imbalance of
sex hormones, there is premenstrual syndrome (PMS). In PMS,
symptoms such as vasomotor nervous symptoms (palpitation, edema),
neuropsychiatric symptoms (headache, dizziness, slight fever,
anxiety, depressive state, insomnia, sleepiness, stress), motor
organ symptoms (lumbago, arthralgia), digestive organ symptoms
(constipation, diarrhea, abdominalgia, hyperphagia, thirst),
systemic symptoms (fatigue), skin symptoms (acne) and the like
appear. A severity of these symptoms is significantly different
among individuals, but when symptom is severe, this hinders work
and daily life in some cases.
[0006] Previously, as response to various symptoms in a menopausal
stage, administration of a sexual steroidal hormone (hormone
replacement therapy (HRT)) has been performed (Non-Patent Document
3).
[0007] However, in treatment in which estrogen is administered for
the menopausal disorder of females, not only there is a report that
an onset risk of breast cancer, venous thromboembolism, cerebral
stroke, ischemic cardiac disease and the like is increased, but
also troublesome administration control and patient management are
required, and a variety of demerits rather than usefulness of
improvement in menopausal symptoms become a problem.
[0008] Further, for the menopausal disorder of males, hormone
replacement therapy with an androgen preparation comprising
testosterone has been spread, but as the side effect, exacerbation
of prostatic cancer, urination disorder, liver function failure,
retention of fluid, exacerbation of sleep apnea syndrome and the
like become a problem.
[0009] In such background, so-called phytoestrogen such as a plant
containing an estrange-like acting substance which is safer and
easily used, and an extract and a fraction obtained therefrom have
been used. As the phytoestrogen, for example, soybean isoflavones
(Patent Document 1, Patent Document 2), black cohosh (Patent
Document 3), Eucommia ulmoides (Patent Document 4), root of
Pueraria radix, Smilax rhizoma, Rehmanniae radix (Patent Document
5) and the like have been reported. Particularly, regarding
isoflavone, equol which is a metabolite is said to be an active
ingredient, and enteric bacteria are involved in conversion to
equol in a body and an individual difference is great in
production, hence, an equol producing composition in which a
microorganism having the ability to produce equol is added to
daidzeins has also been disclosed (Patent Document 6). In these
phytoestrogens, it is said that the effect is relatively mild, and
there is little side effect, but a possibility of an onset risk of
breast cancer or uterine cancer is not denied, and upon ingestion
of a supplement or the like, attention is attracted.
[0010] Further, an aromatase activator which exerts the effect of
preventing, improving or treating each symptom appearing due to
decrease in estrogen by activating aromatase being an enzyme of
converting androgen to estrogen (Patent Document 7), and an
aromatase inhibitor which exerts the effect of preventing/treating
symptom due to decrease in a male hormone or breast cancer in
postmenopausal females by inhibiting aromatase (Patent Document 8)
are also disclosed.
[0011] On the other hand, as a material for alleviating symptom at
a menopausal stage without exhibiting the hormone-like action, a
cacao bean extract-containing product (Patent Document 9),
Tetragonolobus plant and an extract thereof (Patent Document 10), a
rhizome portion powder or an extract of Dioscoreacea plant
containing diosgenin as an active ingredient (Patent Document 11),
a rhizome and an extract of Vitis plant, Japanese knotweed (Patent
Document 12) and the like have been disclosed.
[0012] Meanwhile, it has been found out in an arteriosclerosis
model animal that among phytosterols, a compound having a
cyclolanostane skeleton and a compound having a lophenol skeleton
have the action of reducing the amount of lipid peroxide in blood,
and have the action of suppressing the number of the plaque
formation of thoracic aorta, and use as an antioxidant has been
proposed (Patent Document 13). Further, a sterol obtained from a
plant-derived sterol ester is known to have the pharmacological
action such as the autonomic nerve activating action, the
lipid-lowering action, the platelet aggregation suppressing action,
the cerebral function activating action and the like, and a fat
emulsion containing the sterol which can greatly exert these
pharmacological actions has been disclosed (Patent Document 14).
However, a relationship of a compound having a cyclolanostane
skeleton or a compound having a lophenol skeleton with symptom due
to disturbance in the sex hormone balance has not been reported at
all.
PRIOR ART DOCUMENTS
Non-Patent Documents
[0013] Non-Patent Document 1: Riddle of Hormone Development--Short
Cut for Understanding Environmental Hormones, Yasunobu EGUCHI,
Ishiyaku Pub, Inc. p. 2-5, 2002 [0014] Non-Patent Document 2:
Hormone and Medical Practice, Igakunosekaisha, Inc., Vol. 57, p.
949-954, 2009 [0015] Non-Patent Document 3: Base and Medical
Practice of Antiaging Medicine, edited by Japanese Society of
Anti-Aging Medicine, Medical Specialist Leader Qualification
Committee, p. 214-224, 2006 [0016] Non-Patent Document 4: Basic
Aging Study, Japan Society for Biomedical Gerontology, Vol. 34, p.
13-17, 2010
PATENT DOCUMENTS
[0016] [0017] Patent Document 1: JP-A No. 2007-186483 [0018] Patent
Document 2: JP-A No. 2005-229855 [0019] Patent Document 3: Japanese
Patent No. 4210190 [0020] Patent Document 4: JP-T No. 2012-77012
[0021] Patent Document 5: Japanese Patent No. 4892856 [0022] Patent
Document 6: JP-A No. 2009-232712 [0023] Patent Document 7: JP-A No.
2012-171933 [0024] Patent Document 8: JP-A No. 2014-189539 [0025]
Patent Document 9: JP-A No. 2001-69946 [0026] Patent Document 10:
JP-A No. 2012-31146 [0027] Patent Document 11: JP-A No. 2007-16013
[0028] Patent Document 12: Japanese Patent No. 4768105 [0029]
Patent Document 13: WO 2010/058795 [0030] Patent Document 14: JP-A
No. 4-91026
DISCLOSURE OF THE INVENTION
Problems to be Solved by the Invention
[0031] It has been confirmed that the materials disclosed in Patent
Documents 9 to 12 exhibit the effect of improving symptoms which
can occur at a menopausal stage, as one example of the effect on
symptoms caused by an imbalance of sex hormones, but that effect is
only on a part of symptoms, and the direct effect on symptoms
caused by an imbalance of sex hormones has not been confirmed.
Further, the effect confirmed in Patent Document 14 is only the
platelet aggregation suppressing action, and there was no
description regarding other pharmacological actions.
[0032] Then, an object of the present invention is to provide a
functional material which can safely be ingested daily and can
effectively prevent or improve symptoms caused by disturbance in
the sex hormone balance, with as little pain as possible, and a
agent utilizing this.
Means to Solve the Problems
[0033] The first invention which solves the above-mentioned
problems is an agent for preventing or improving symptoms caused by
an imbalance of sex hormones, which contains, as an active
ingredient, a compound selected from the group consisting of
lophenol compounds and cyclolanostane compounds (hereinafter,
referred to also as "agent of the present invention").
[0034] In a preferable embodiment of the present invention, the
agent comprises the above-described compound at a total amount of
0.00001% by mass or more.
[0035] In a preferable embodiment of the present invention, the
above-described compound is selected from the group consisting of
4-methylcholest-7-en-3-ol, 4-methylergost-7-en-3-ol,
4-methylstigmast-7-en-3-ol, 9,19-cyclolanostan-3-ol and
24-methylene-9,19-cyclolanostan-3-ol.
[0036] The agent of the present invention is effective in
preventing or improving symptoms caused by an imbalance of female
hormones.
[0037] In a preferable embodiment of the present invention, the
agent of the present invention is an agent which prevents or
improves the symptom not through the sex hormone-like action.
[0038] The symptoms caused by an imbalance of sex hormones include
bone dysbolism and neuropsychiatric symptoms, and the
above-described compound or the composition containing this is
particularly effective in preventing or improving these
symptoms.
[0039] Further, the second invention for solving the
above-mentioned problem is use of a compound selected from the
group consisting of lophenol compounds and cyclolanostane compounds
in the manufacture of an agent for preventing or improving symptoms
caused by an imbalance of sex hormones, and a preferable embodiment
of the compound is as described above.
[0040] Further, the second invention includes the following
embodiment.
[0041] Use of a composition containing a compound selected from the
group consisting of lophenol compounds and cyclolanostane compounds
at a total amount of 0.00001% by mass or more in the manufacture of
an agent for preventing or improving symptoms caused by an
imbalance of sex hormones.
[0042] In a preferable embodiment of the present invention, the
agent is for preventing or improving symptoms caused by an
imbalance of female hormones.
[0043] In a preferable embodiment of the present invention, the
agent is for preventing or improving the above-described symptoms
not through the sex hormone-like action.
[0044] In a preferable embodiment of the present invention, the
agent is for preventing or improving bone dysbolism or
neuropsychiatric symptoms.
[0045] Further, the third invention for solving the above-mentioned
problem is a compound selected from the group consisting of
lophenol compounds and cyclolanostane compounds, which is used for
preventing or improving symptoms caused by an imbalance of sex
hormones, and a preferable embodiment of the compound is as
described above.
[0046] In a preferable embodiment of the present invention, the
symptoms caused by an imbalance of sex hormones are symptoms caused
by an imbalance of female hormones.
[0047] In a preferable embodiment of the present invention, the
above-described compound is for preventing or improving the
symptoms not through the sex hormone-like action.
[0048] In a preferable embodiment of the present invention, the
above-described symptoms caused by an imbalance of sex hormones are
bone dysbolism and neuropsychiatric symptoms.
[0049] Further, the fourth invention for solving the
above-mentioned problem is a composition containing a compound
selected from the group consisting of lophenol compounds and
cyclolanostane compounds at a total amount of 0.00001% by mass or
more, which is used for preventing or improving symptoms caused by
an imbalance of sex hormones, and a preferable embodiment of the
compound is as described above.
[0050] In the fourth invention, the above-described composition is
preferably food or drink.
[0051] In a preferable embodiment of the present invention, the
symptoms caused by an imbalance of sex hormones are symptoms caused
by an imbalance of female hormones.
[0052] In a preferable embodiment of the present invention, the
above-described composition is for preventing or improving the
symptoms not through the sex hormone-like action.
[0053] In a preferable embodiment of the present invention, the
symptoms caused by an imbalance of sex hormones are bone dysbolism
and neuropsychiatric symptom.
[0054] Further, the fifth invention for solving the above-mentioned
problem is a method for preventing or improving symptoms caused by
an imbalance of sex hormones, comprising administering a compound
selected from the group consisting of lophenol compounds and
cyclolanostane compounds to a subject having symptoms caused by an
imbalance of sex hormones, and a preferable embodiment of the
compound is as described above.
[0055] Further, the fifth invention has the following preferable
embodiment:
[0056] The composition containing a compound selected from the
group consisting of lophenol compounds and cyclolanostane compounds
at a total amount of 0.00001% by mass or more is administered to
the above-described subject.
[0057] In a preferable embodiment of the present invention, the
above-described symptoms caused by an imbalance of sex hormones are
symptoms caused by an imbalance of female hormones.
[0058] In a preferable embodiment of the present invention, the
prevention or the improvement is conducted not through the sex
hormone-like action.
[0059] In a preferable embodiment of the present invention, the
above-described symptoms caused by an imbalance of sex hormones are
bone dysbolism and neuropsychiatric symptoms.
Effect of the Invention
[0060] The agent of the present invention can be safely ingested,
and efficiently prevents or improves symptoms caused by an
imbalance of sex hormones. Particularly, the agent of the present
invention is effective in preventing or improving symptoms caused
by an imbalance of female hormones. Further, the agent of the
present invention is effective in preventing or improving,
particularly, bone dysbolism or neuropsychiatric symptoms being a
symptom caused by an imbalance of sex hormones.
DESCRIPTION OF EMBODIMENTS
[0061] Then, preferable embodiments of the present invention will
be illustrated in detail. The present invention is not limited to
the following preferable embodiments, and can be freely modified in
the scope of the present invention. In addition, percentage in the
present description is indication by mass, unless otherwise
indicated. Further, the "preventing or improving agent" of the
present invention (in the present description, simply referred to
as "agent of the present invention" in some cases) has the same
meaning as that of the "preventing or treating agent" of the
present invention, and every case is included in definition of the
agent of the present invention. Further, in the present
description, "improvement" is the concept including
"treatment".
[0062] The agent of the present invention contains, as an active
ingredient, a compound selected from the group consisting of
lophenol compounds (Compound 1) and cyclolanostane compounds
(Compound 2). Compound 1 and Compound 2 are represented by the
following general formula (1) and the general formula (2),
respectively.
##STR00001##
[0063] In the general formula (1), R1 is an alkyl group or an
alkenyl group including one or two double bonds, which is straight
or branched chain having 5 to 16 carbon atoms. The alkyl or alkenyl
group may be a substituted alkyl or alkenyl group, in which one or
two hydrogen atoms are substituted with a hydroxyl group and/or a
carbonyl group.
[0064] R2 and R3 each are independently a hydrogen atom or an alkyl
group having 1 to 3 carbon atoms. Herein, as the alkyl group having
1 to 3 carbon atoms, a methyl group, an ethyl group and the like
are preferable, and a methyl group is particularly preferable. The
alkyl group may be a substituted alkyl group in which at least one
hydrogen atom is substituted with a hydroxyl group and/or a
carbonyl group.
--CH.sub.2--OH
--CH.sub.2--COOH
--CH.sub.2--CH.sub.2--OH
--CH.sub.2--CH.sub.2--COOH
--CH(OH)--CH.sub.3
--CH(COOH)--CH.sub.3 [Chemical formula 2]
[0065] R4 forms C.dbd.O with a carbon atom constituting the ring,
or is --OH or --OCOCH.sub.3.
[0066] In the general formula (1), R1 is preferably any of groups
represented by the following formulae.
--CH.sub.2--CH.sub.2--CH(CH.sub.2--CH.sub.2)--CH(CH.sub.2).sub.2
--CH.sub.2--CH.sub.2--CH.dbd.C(CH.sub.3).sub.2
--CH.sub.2--CH.dbd.C(CH.sub.2)--CH(CH.sub.2).sub.2
--CH.sub.2--CH.sub.2--C(.dbd.CH--CH.sub.3)--CH(CH.sub.3).sub.2
--CH.sub.2--CH.sub.2--CH(Ra)=C(CH.sub.2)Rb [Chemical formula 3]
(wherein Ra and Rb are any of a hydrogen atom, a hydroxyl group and
a methyl group)
--CH.sub.2--CH.sub.2--CH(Rc)-CH(CH.sub.3)Rd
(wherein Rc and Rd are any of a hydrogen atom, a hydroxyl group and
a methyl group)
[0067] In the general formula (1), it is preferable that one of R2
and R3 is a hydrogen atom, and the other is a methyl group, and it
is preferable that R4 is a hydroxy group.
[0068] Compound 1 includes preferably 4-methylcholest-7-en-3-ol,
4-methylergost-7-en-3-ol and 4-methylstigmast-7-en-3-ol. Respective
compounds have structures represented by the following formulae,
respectively.
##STR00002##
[0069] Compound 1 can be chemically manufactured in accordance with
the known manufacturing processes.
[0070] Compound 1 can be synthesized, for example, in accordance
with supplement data described in Vitali Matyash et al., PLOS
BIOLOGY, Volume 2, Issue 10, e280, 2004.
[0071] Further, it is known that Compound 1 is contained in plants,
and Compound 1 can be manufactured in accordance with the known
process for manufacturing lophenol (Biochemistry Experimental
Method 24, Fat Lipid Metabolism Experimental Method, authored by
Akihiro YAMADA, Gakkai Shuppan Center, p. 174, 1989).
[0072] For example, Compound 1 can be extracted from plants which
are known to contain Compound 1, using a method such as a hot water
extraction method, an organic solvent extraction method, a
supercritical extraction method and a subcritical extraction method
(see, e.g., Japanese Patent No. 3905913). Compound 1 can be
extracted, for example, from plants belonging to family Liliaceae,
family Leguminosae, family Gramineae, family Solanaceae and family
Musaseae.
[0073] The molecular weight and the structure of Compound 1
manufactured as described above can be determined or confirmed by a
mass spectrometry (MS), a nuclear magnetic resonance spectral (NMR)
method.
[0074] Further, Compound 1 may be a pharmaceutically acceptable
salt. The pharmaceutically acceptable salt includes both metal
salts (inorganic salts) and organic salts, and as a list of them,
that described in "Remington's Pharmaceutical Sciences, 17th
edition, 1985, p. 1418" is exemplified.
[0075] Specifically, inorganic salts such as a hydrochloride, a
sulfate, a phosphate, a diphosphate, and a hydrobromide, and
organic salts such as a malate, a maleate, a fumarate, a tartrate,
a succinate, a citrate, an acetate, a lactate, a methanesulfonate,
a p-toluenesulfonate, a pamoate, a salicylate, and a stearate are
included without limitation.
[0076] Meanwhile, Compound 1 may be a salt with a metal such as
sodium, potassium, calcium, magnesium and aluminum, or a salt with
an amino acid such as lysine. Moreover, there may also be used a
solvate such as a hydrate of the compounds or pharmaceutically
acceptable salts thereof.
##STR00003##
[0077] In the general formula (2), R5 is an alkyl group or an
alkenyl group including one or two double bonds, which is straight
or branched chain having 6 to 8 carbon atoms. The alkyl or alkenyl
group may be a substituted alkyl or alkenyl group in which one or
two hydrogen atoms are substituted with a hydroxyl group and/or a
carbonyl group.
[0078] R6 and R7 each are independently a hydrogen atom or a methyl
group. R8 forms C.dbd.O with a carbon atom constituting the ring,
or is any of the following formulae.
##STR00004##
[0079] In the general formula (2), R5 is preferably any of groups
represented by the following formulae.
--CH.sub.2--CH.sub.2--CH.sub.2--CH(CH.sub.3).sub.2
--CH.sub.2--CH.sub.2--CHRe--C(CH.sub.3).sub.2Rf [Chemical formula
9]
(Re is a hydrogen atom, a hydroxyl group or a methyl group, and Rf
is a hydrogen atom or a hydroxyl group)
--CH.sub.2--CH.sub.2--CH(CH.sub.2--CH.sub.3)--CH(CH.sub.3).sub.2
--CH.sub.2--CH.sub.2--CHRg--C(CH.sub.3).dbd.CH.sub.2
(Rg is a hydrogen atom, a hydroxyl group or a methyl group)
--CH--CH.sub.2--C(.dbd.O)--CH(CH.sub.3).dbd.CH.sub.2
--CH.sub.2--CH.sub.2--C(.dbd.CH.sub.2)--CH(CH.sub.3).sub.3
--CH.sub.2--CH.sub.2--CH.dbd.C(CH.sub.3).sub.2
--CH.sub.2--CH.sub.2.dbd.C(CH.sub.3)--CH(CH.sub.3).sub.2
--CH.sub.2--CH.sub.2--C(.dbd.CH--CH.sub.3)--CH(CH.sub.3).sub.3
[0080] Further, in the general formula (2), it is preferable that
one of R6 and R7 is a hydrogen atom, and the other is a methyl
group, and it is preferable that R8 is a hydroxy group.
[0081] Compound 2 includes preferably 9,19-cyclolanostan-3-ol and
24-methylene-9,19-cyclolanostan-3-ol. Respective compounds have
structures represented by the following formulae, respectively.
##STR00005##
[0082] Compound 2 can be chemically manufactured in accordance with
the known manufacturing processes. For example,
24-methylene-9,19-cyclolanostan-3-ol (trivial name:
24-methylenecycloartanol) can be manufactured by the methods
disclosed in JP-A No. 57-018617 and WO 2012/023599 (method of
synthesis from .gamma.-oryzanol). Alternatively, Compound 2 can be
manufactured using a hydrolysate of cycloartenol ferulate as a
starting substance, by the method disclosed in JP-A No.
2003-277269.
##STR00006##
[0083] Further, Compound 2 is also known to be contained in a plant
belonging to family Liliaceae, family Leguminosae, family
Gramineae, family Solanaceae, or family Musaseae (see
[Phytochemistry, USA, 1977, vol. 16, pp. 140-141], [Handbook of
phytochemical constituents of GRAS herbs and other economic plants,
1992, USA, CRC Press] or [Hager's Handbuch der Pharmazeutischen
Praxis, vol. 2-6, 1969-1979, Deutschland, Springer Verlag,
Berlin]). Hence, Compound 2 can be extracted from these plants
using the known methods such as an organic solvent extraction
method or a hot water extraction method (see, e.g., Japanese Patent
No. 3924310). It is preferable that Compound 2 is extracted, for
example, from plants of Liliaceae Aloe.
[0084] The molecular weight and the structure of the compound
manufactured as described above can be determined or confirmed, for
example, by mass spectrometry (MS) and nuclear magnetic resonance
spectrometry (NMR).
[0085] Further, Compound 2 may be a pharmaceutically acceptable
salt. Such a salt is as exemplified concerning Compound 1.
[0086] The agent of the present invention contains, as an active
ingredient, a compound selected from the group consisting of
Compound 1 and Compound 2. The compound may be one kind, that is,
either of Compound 1 or Compound 2 alone, or may be a mixture of
Compound 1 and Compound 2.
[0087] When Compound 1 or Compound 2 is used alone, either Compound
1 (mainly, 4-methylcholest-7-en-3-ol, 4-methylergost-7-en-3-ol, or
4-methylstigmast-7-en-3-ol) or Compound 2 (mainly,
9,19-cyclolanostan-3-ol or 24-methylene-9,19-cyclolanostan-3-ol) is
preferable.
[0088] Among them, 4-methylcholest-7-en-3-ol is particularly
preferable as Compound 1, and 9,19-cyclolanostan-3-ol is
particularly preferable as Compound 2, from a view point of
physical properties such as solubility which are considered when
used as an active ingredient of the agent.
[0089] Further, when Compound 1 and Compound 2 are compared,
Compound 1 (mainly, 4-methylcholest-7-en-3-ol,
4-methylergost-7-en-3-ol or 4-methylstigmast-7-en-3-ol) is more
preferable.
[0090] Further, for each of Compound 1 or Compound 2, one kind of a
compound may be used, or a plurality of compounds may be used by
mixing them.
[0091] The agent of the present invention contains, as an active
ingredient, preferably a compound selected from the group
consisting of 4-methylcholest-7-en-3-ol, 4-methylergost-7-en-3-ol,
4-methylestigmast-7-en-3-ol, 9,19-cyclolanostan-3-ol and
24-methylene-9,19-cyclolanostan-3-ol.
[0092] The content of the compound in the agent of the present
invention can be appropriately selected depending on symptoms or
the like, and the total amount is preferably at least 0.00001% by
mass, more preferably at least 0.0001% by mass, further preferably
at least 0.0005% by mass, and particularly preferably at least
0.001% by mass. Further, the upper limit of the amount in the agent
of the present invention is not particularly limited, but as the
total amount, 90% by mass or less, preferably 70% by mass or less
and more preferably 50% by mass or less are exemplified.
[0093] When both Compound 1 and Compound 2 are combined (mixture of
Compound 1 and Compound 2), the range of the mass ratio of Compound
1 and Compound 2 includes, for example, the following:
[0094] Compound 1:Compound 2 is preferably 5:1 to 1:5, further
preferably 3:1 to 1:3, and particularly preferably 2:1 to 1:2.
[0095] For example, as an example in which the compound is
contained in a natural plant, it is known that Compound 1 (mainly,
4-methylcholest-7-en-3-ol, 4-methylergost-7-en-3-ol and
4-methylstigmast-7-en-3-ol) and Compound 2 (mainly,
9,19-cyclolanostan-3-ol and 24-methylene-9,19-cyclolanostan-3-ol)
are contained in aloe vera.
[0096] For this reason, using aloe vera as a raw material, any of
4-methylcholest-7-en-3-ol, 4-methylergost-7-en-3-ol or
4-methylstigmast-7-en-3-ol (Compound 1) or any of
9,19-cyclolanostan-3-ol, or 24-methylene-9,19-cyclolanostan-3-ol
(Compound 2) can be purified, respectively, to obtain a mixture
containing Compound 1:Compound 2 at 5:1 to 1:5, preferably 3:1 to
1:3, and particularly preferably 2:1 to 1:2. Thus obtained mixture
is suitable as an active ingredient of the agent of the present
invention.
[0097] The agent of the present invention can be used in an aspect
of a medicament.
[0098] The agent of the present invention can be orally or
parenterally administered to a mammal including a human.
[0099] In the present invention, as symptoms caused by an imbalance
of sex hormones, symptoms caused by a menopausal disorder, a
postmenopausal disorder, a juvenile menopausal disorder, a
menstrual disorder, dysmenorrhea, a sleep disorder and the like in
addition to bone dysbolism, neuropsychiatric symptoms and the like
can also be exemplified.
[0100] For example, symptoms caused by a menopausal disorder
include vasomotor nervous symptoms (glow, hot flash, sweating,
coldness, palpitation, short breath, edema), neuropsychiatric
symptoms (headache, dizziness, insomnia, anxiety, irritation,
depressive state, tinnitus, deafness, lightheadedness, stress,
dyspnea), motor organ symptoms (lumbago, shoulder stiffness,
arthralgia, myalgia, bone dysbolism), digestive organ symptoms
(anorexia, nausea, diarrhea, constipation, thirst, dull feeling in
the stomach, heartburn), systemic symptoms (malaise), urinary
symptoms (pollakiuria, residual urine, micturition pain, reduction
in urinary stream), reproduction organ symptoms (menstrual
disorder, reduction in sexual desire), sensory symptoms (numbness,
desensitization, hypersensitivity, low vision), skin symptoms (skin
drying, itching, wrinkle, dullness) and sleep disorders (sleep
apnea syndrome, etc.). Other examples include rheumatoid arthritis,
goiter and the like. These symptoms are seen at a menopausal stage
in males and females.
[0101] Further, symptoms caused by a postmenopausal disorder
include hypertension, diabetes, obesity, osteoporosis, unidentified
complaint, dementia and the like.
[0102] Further, symptoms of premenstrual syndrome include vasomotor
nervous symptoms (palpitation, edema), neuropsychiatric symptoms
(headache, dizziness, slight fever, anxiety, depressive state,
insomnia, sleepiness, stress), motor organ symptoms (lumbago,
arthralgia), digestive organ symptoms (constipation, diarrhea,
abdominalgia, hyperphagia, thirst), systemic symptoms (fatigue),
and skin symptoms (acne).
[0103] Inter alia, the agent of the present invention is effective
for symptoms due to an imbalance of female hormones. As the female
hormone, estrogen including esteron, estradiol and estriol
(follicular hormone), and progesterone (corpus luteum hormone) are
known. Among them, estrogen has its receptors in a variety of
tissues, and acts on the reproduction function, the cardiac blood
vessel, the skeleton, the skin and the like. For this reason, the
agent of the present invention is particularly effective in
symptoms caused by an imbalance of female hormones due to
deficiency of estrogen. Such symptoms include bone dysbolism,
neuropsychiatric symptoms and the like. Further, symptoms caused by
a menopausal disorder, a postmenopausal disorder, a juvenile
menopausal disorder, a menstrual disorder, dysmenorrhea,
premenstrual syndrome and the like can also be exemplified.
[0104] In addition, symptoms caused by a female menopausal disorder
include vasomotor nervous symptoms (glow, hot flash, sweating,
coldness, palpitation, short breath, edema), neuropsychiatric
symptoms (headache, dizziness, insomnia, anxiety, irritation,
depressive state, tinnitus, deafness, lightheadedness, stress),
motor organ symptoms (lumbago, shoulder stiffness, arthralgia,
myalgia, bone dysbolism), digestive organ symptoms (anorexia,
nausea, diarrhea, constipation, thirst, dull feeling in the
stomach, heartburn), systemic symptoms (malaise), urinary symptoms
(pollakiuria, residual urine, micturition pain), reproduction organ
symptoms (menstrual disorder, reduction in sexual desire), sensory
symptoms (numbness, desensitization, hypersensitivity, low vision)
and skin symptoms (skin drying, itching, wrinkle, dullness). Other
examples include hypertension, diabetes, obesity, osteoporosis,
autonomic ataxia, unidentified complaint, dementia and the like
which are said to be developed after menopause. Symptoms caused by
a postmenopausal disorder and premenstrual syndrome are as stated
above.
[0105] The agent of the present invention has the effect of
preventing or improving the symptoms not through the sex
hormone-like action. That is, the agent of the present invention
influences symptoms caused by an imbalance of sex hormones, and
exerts the action of preventing or improving the symptoms.
Accordingly, the present invention can provide the excellent agent
in a point that a difference in sex of a subject influences little
difference in the effect of preventing or improving symptoms, and
in a point of safety, for example.
[0106] The agent of the present invention is effective in
preventing or improving one or a plurality of symptoms among the
above-mentioned symptoms. Further, as shown in Examples described
later, the agent of the present invention is particularly useful in
that it has the effect of preventing or improving a plurality of
symptoms among these symptoms.
[0107] The effect of the agent of the present invention can be
confirmed using an ovariectomized mouse (OVX mouse) which is a
model animal triggering an imbalance of sex hormones.
[0108] Since the OVX mouse triggers an imbalance of sex hormones by
deficiency in sexual hormones, it develops a variety of symptoms
due to this. In the OVX mouse, due to an imbalance of sex hormones,
for example, bone dysbolism (reduction in the bone-specific
alkaline phosphatase concentration in blood) or neuropsychiatric
symptoms (increase in the corticosterone concentration in blood)
are observed (EXPERIMENTAL AND THERAPEUTIC MEDICINE, v.8, p.
957-967, 2014, Metab Brain Dis, (2013), v. 28, p. 77-84).
[0109] That is, when the agent is administered to the OVX mouse, if
increase in the bone-specific alkaline phosphatase concentration in
blood or reduction in the corticosterone concentration in blood is
confirmed, it is supported that there is obtained the effect of
preventing or improving symptoms due to an imbalance of sex
hormones, including bone dysbolism and neuropsychiatric symptoms,
by the administered agent.
[0110] The agent of the present invention exhibits the remarkable
effect on prevention or improvement of motor organ symptoms such as
bone dysbolism and the like and neuropsychiatric symptoms, among
symptoms due to an imbalance of sex hormones. The effect of
preventing or improving bone dysbolism includes the osteogenesis
promoting effect. Further, the effect of preventing or improving
neuropsychiatric symptom includes the effect of preventing or
improving symptoms caused by ultraviolet stress.
[0111] The agent of the present invention is also effective in
preventing or improving reduction in the bone density or
osteoporosis due to bone dysbolism, or unidentified complaint which
can be mainly caused by neuropsychiatric symptoms. Further, the
agent of the present invention is also effective in preventing or
improving these plural symptoms.
[0112] The form of the agent of the present invention is not
particularly limited, and can be appropriately selected depending
on a usage. Specifically, tablets, pills, powders, liquids,
suspensions, emulsions, granules, capsules, syrups, suppositories,
injections, ointments, patches, eye drops, nose drops and the like
can be exemplified.
[0113] The administration term of the agent of the present
invention is not particularly limited, and can be appropriately
selected depending on a target disease. Further, it is preferable
that the dose is determined depending on a dosage form, dose
regimen, age and sex of a patient, other conditions, degree of
symptom and the like.
[0114] The dose of the agent of the present invention is
appropriately selected depending on a dose regimen, age and sex of
a patient, degree of a disease, other conditions and the like.
Usually, as the dose in terms of an amount of an active ingredient,
a standard is preferably in the range of 0.0001 to 100 mg/day, more
preferably 0.001 to 50 mg/day, and particularly preferably 0.01 to
10 mg/day.
[0115] The agent of the present invention may contain an additive
which is generally used in a medicament. The additive includes an
excipient, a binder, a disintegrating agent, a lubricant, a
stabilizer, a flavoring agent, a diluent, a surfactant, a solvent
for injection and the like.
[0116] The agent of the present invention can be manufactured by
blending the above-described compound as an active ingredient into
a medicament carrier. The agent of the present invention can be
manufactured, for example, by formulating the compound together
with the additive described above into a preparation.
[0117] Alternatively, the agent of the present invention can also
be manufactured by formulating a compound obtained by extraction
using hot water or various solvents, supercritical extraction, or
subcritical extraction, using the known plant containing the
compound or the like as a raw material, together with the additives
into preparations.
[0118] Particularly, the agent of the present invention comprising
Compound 1 and Compound 2 in the specific range of the mass ratio
can be manufactured by mixing respective compounds in the
above-mentioned range of the mass ratio. Alternatively, such a
medicament can also be manufactured by a method of extraction using
various solvents, supercritical extraction, subcritical extraction
or the like, using the known plant containing a mixture containing
Compound 1 and Compound 2 or the like as a raw material.
[0119] In the agent of the present invention, the above-described
compound functions as an active ingredient, and has the action of
preventing or improving symptoms caused by an imbalance of sex
hormones. Particularly, the agent is effective for symptoms due to
an imbalance of female hormones. Further, inter alia, the agent
exerts the remarkable preventing or improving effect on bone
dysbolism or neuropsychiatric symptoms.
[0120] The agent of the present invention can also be processed
into food or drink by mixing it with raw materials which can be
used in food or drink. In the present invention, "food or drink"
includes feed which is ingested by animals other than a human, in
addition to food or drink which is ingested by a human.
[0121] When food or drink is manufactured, the amount of the
compound in them is, as the total amount, preferably at least
0.00001% by mass, more preferably at least 0.0001% by mass, further
preferably at least 0.0005% by mass, and particularly preferably at
least 0.001% by mass. Further, the upper limit of the amount in
food or drink of the present invention is not particularly limited,
but as the total amount, 90% by mass or less, preferably 70% by
mass or less, and more preferably 50% by mass or less are
exemplified.
[0122] Further, the amount of the compound in food or drink can
also be made to be a suitable amount for the compound to be
ingested in the range of preferably 0.0001 to 100 mg/day, more
preferably 0.001 to 50 mg/day, and particularly preferably 0.01 to
10 mg/day, as expressed by the total amount, depending on the form
thereof. Accordingly, one of preferable forms of food or drink of
the present invention is used so that the compound is ingested at
preferably 0.0001 to 100 mg/day, more preferably 0.001 to 50
mg/day, and particularly preferably 0.01 to 10 mg/day, as expressed
by the total amount.
[0123] The food or drink is preferably a health functional food or
drink. The "health functional food or drink" means a food or drink
which directly or indirectly indicates the effect of preventing a
disease, or the effect of reducing an onset risk of a disease, and
a food or drink which was notified at Consumer Affairs Agency as
indicating the functionality on a merchandise package based on
scientific basis under business operator's responsibility. Examples
thereof include foods or drinks which are currently sold as a food
for specified health use, a food with function claims, a health
supplement or the like in Japan.
[0124] The form of food or drink is not particularly limited, but
drinks such as a soft drink, a carbonated drink, a nutritional
drink, a fruit juice drink, a lactic acid bacteria beverage and the
like (including concentrated original liquid and powder for
preparation of these drinks) are particularly preferable from a
view point that the compound is effectively ingested.
[0125] Further, it is preferable that the form of functional food
or drink is a granule, tablet or liquid supplement, from a view
point that a person who ingests it can easily grasp the ingestion
amount of an active ingredient.
[0126] Further, it is also preferable that such functional food or
drink is in a form with an indication of use of "for preventing or
improving symptoms caused by an imbalance of sex hormones", "for
restoring the sex hormone balance", "for preventing or improving
bone dysbolism", or "for preventing or improving neuropsychiatric
symptom" attached thereto. That is, it is preferable that the food
or drink of the present invention is sold, for example, as food or
drink for preventing or improving symptoms caused by an imbalance
of sex hormones, containing a compound selected from the group
consisting of Compound 1 and Compound 2 as an active ingredient,
with use of "for preventing or improving symptoms caused by an
imbalance of sex hormones" attached thereto.
[0127] The "indication" includes all indications having the
function of informing consumers of the use. That is, indications
which can evoke or analogize the use all correspond to the
"indication", irrespective of an object of an indication, the
content of an indication, a subject to be indicated, a medium and
the like with which an indication is performed.
[0128] Further, the "with an indication attached thereto" refers to
that an indication act of making consumers recognize the indication
associated with food or drink (products) exists.
[0129] It is preferable that an indication act is such that
consumers can directly recognize the use. Specifically, an act of
describing the use on merchandise related to the food or drink of
the present invention or a package of merchandise, and an act of
describing the use on advertisement regarding merchandise, a price
list or transaction documents (including those provided by
electromagnetic method) are exemplified.
[0130] On the other hand, it is preferable that the content to be
indicated (indication content) is an indication approved by
administration or the like (for example, indication which received
approval based on various institutions provided by administration,
and is performed in an aspect based on such approval).
[0131] For example, indications of a health food, functional food
or drink, an enteral nutritive food, a food for special dietary
uses, a food with health claims, a special health food, a food with
nutrient function claims, a food with function claims, a quasi-drug
and the like can be exemplified. Particularly, indication approved
by Consumer Affairs Agency, for example, indications approved by
special health food institution, or institutions similar thereto
can be exemplified. As an example of the latter, an indication as a
special health food, an indication as a conditional special health
food, an indication to the effect that a structure or the function
of a body is influenced, a disease risk reducing indication and the
like can be exemplified, more particularly, an indication as a
special health food provided in Health Promotion Act, Enforcement
Regulation (2003 Apr. 30 Japanese Ordinance of the Ministry of
Health, Labour and Welfare No. 86) (particularly, indication of use
of health), and indications similar thereto can be mentioned as a
typical example.
[0132] Phrase indicating the use is not limited to phrase of "for
preventing or improving symptoms caused by an imbalance of sex
hormones", "restoring the sex hormone balance", "for preventing or
improving bone dysbolism", or "for preventing or improving
neuropsychiatric symptom", and it goes without saying that other
phrases are included in the scope of the present invention as far
as they are phrase indicating the action or the effect of
alleviating or improving various symptoms caused by an imbalance of
sex hormones.
[0133] Further, it is also preferable that the food or drink of the
present invention includes an indication of the active ingredient,
and further, an indication showing the relevancy between the use
and the active ingredient, in addition to the indication of
use.
[0134] The food or drink can be manufactured by blending a compound
selected from Compound 1 and Compound 2 as an active ingredient.
The food or drink of the present invention can be manufactured, for
example, by mixing the compound into food or drink raw materials,
followed by processing.
[0135] Alternatively, the food or drink can also be manufactured by
processing an extract obtained by extraction using hot water or
various solvents, supercritical extraction, or subcritical
extraction using the known plant containing the compound or the
like as a raw material, together with food or drink raw
materials.
[0136] Further, when a form of the food or drink is made to be a
granular, tablet-shaped or liquid supplement, it is also preferable
that the compound being an active ingredient together with, for
example, saccharides such as lactulose, maltitol, and lactitol, and
other saccharides, for example, dextrin, starch and the like;
proteins such as gelatin, soybean protein, and corn protein; amino
acids such as alanine, glutamine, and isoleucine; polysaccharides
such as cellulose and gum arabic; fats or oils such as soybean oil,
and medium chain fatty acid triglyceride is formulated into a
preparation.
EXAMPLES
[0137] The present invention will be illustrated in more detail
below by way of Examples, but the present invention is not limited
to the following Examples.
Production Example 1
(Production of Lophenol Compound (Compound 1))
[0138] Mesophyll of aloe vera (transparent gel portion) (100 kg)
was liquefied using a homogenizer, and 100 L of an ethyl
acetate/butanol (3:1) mixed liquid was added thereto, followed by
stirring. After allowing to stand overnight, the ethyl
acetate/butanol mixed liquid and an aqueous layer were separated to
recover the ethyl acetate/butanol mixed liquid. This ethyl
acetate/butanol mixed liquid was concentrated under reduced
pressure. The mass of the recovered ethyl acetate/butanol mixed
liquid extract was 13.5 g.
[0139] A solution obtained by dissolving 13 g of the extract in 1
ml of a chloroform/methanol (1:1) mixed liquid was passed through a
column filled with 400 g of Silica Gel 60 (manufactured by Merck
& Co., Inc.), the extract was adsorbed thereon, then, eluted by
a stepwise gradient method of stepwisely increasing the methanol
concentration using a chloroform/methanol mixed liquid (each mixing
ratio of chloroform:methanol=100:1, 25:1, 10:1, 5:1 and 1:1), and
the eluted liquid was fractionated for every mixed ratio of the
mixed liquid. It was confirmed by normal phase and reverse phase
thin layer chromatography (manufactured by Merck & Co., Inc.,
Silica Gel 60F254 and RP-18F2543) that the lophenol compound of the
present invention exists in a fraction which had been eluted at
chloroform:methanol=25:1, among these fractions.
[0140] The solvent of this fraction was removed, then, the residue
was dissolved in 1 ml of a chloroform/methanol (1:1) mixed liquid,
passed through a column filled with 100 g of Silica Gel 60,
adsorbed onto a column, then, eluted with 1100 ml of a hexane/ethyl
acetate (4:1) mixed liquid. Eluted fractions were collected by 300
ml (Fraction A), 300 ml (Fraction B), and 500 ml (Fraction C) in
this order.
[0141] It was confirmed by normal phase and reverse phase thin
layer chromatography that the lophenol compound being Compound 1 of
the present invention was concentrated in Fraction A, and further
separated with a chloroform/hexane (85:15) mixed liquid using HPLC
equipped with COSMOSIL C18 (manufactured by Nacalai Tesque, Inc.)
to obtain 4-methylcholest-7-en-3-ol, 4-methylergost-7-en-3-ol, and
4-methylstigmast-7-en-3-ol at 1.3 mg, 1.2 mg, and 1 mg,
respectively. The structure of each compound was confirmed by mass
spectrometry (MS) and NMR.
Production Example 2
(Production of Cyclolanostane Compound (Compound 2))
[0142] To 8.0 g of .gamma.-oryzanol (manufactured by Oryza Oil
& Fat Chemical Co., Ltd.) were added 250 ml of distilled water,
50 g of sodium hydroxide, 150 ml of isopropanol, 150 ml of ethanol,
and 150 ml of methanol, and heating refluxing was performed for 2
hours using a mantle heater. After the reaction, the reaction
liquid was added to 1300 ml of water, and the produced white
precipitate was suction-filtered to obtain a solid. In order to
wash the remaining alkali, the resulting residue was suspended in
1000 ml of water, and suction filtration was performed again. This
operation was repeated two times, and the final residue was
lyophilized under reduced pressure to obtain 5.91 g of an oryzanol
hydrolysate. The hydrolysate was purified by HPLC to obtain 2435 mg
of cycloartenol, and 1543 mg of
24-methylene-9,19-cyclolanostan-3-ol (Compound 2).
[0143] Then, using the resulting cycloartenol,
9,19-cyclolanostan-3-ol (Compound 2) was synthesized.
[0144] 302 mg of cycloartenol, 150 ml of isopropanol, and 1.0 g of
a powdery 5% palladium-carrying carbon catalyst were charged, these
were sealed in an autoclave, the interior thereof was replaced with
a nitrogen gas, and a hydrogen gas was introduced while applying a
pressure of 3 kg/cm.sup.2. This was heated while stirring, and at
the time point at which a temperature became 50.degree. C., a
pressure of hydrogen was adjusted at 5 kg/cm.sup.2, and a reaction
was performed for 6 hours while retaining a pressure by
supplementing absorbed hydrogen. The reaction liquid was filtered
to remove the catalyst, concentrated and purified by silica gel
column chromatography (developing solvent: chloroform 100%) to
obtain 275 mg of 9,19-cyclolanostan-3-ol.
Production Example 3
(Preparation of Sample Containing Mixture of Lophenol Compound and
Cyclolanostane Compound Added Thereto)
[0145] Using 4-methylcholest-7-en-3-ol, 4-methylergost-7-en-3-ol
and 4-methylstigmast-7-en-3-ol which had been obtained by
Production Example 1, and 9,19-cyclolanostan-3-ol and
24-methylene-9,19-cyclolanostan-3-ol which had been obtained by
Production Example 2, a mixture in which the mass ratio of the
lophenol compound and the cyclolanostane compound was lophenol
compound (Compound 1):cyclolanostane compound (Compound 2)=1:1 was
obtained.
[0146] Using carboxymethylcellulose (CMC: manufactured by DKS Co.,
Ltd.), a mixture of the lophenol compound and the cyclolanostane
compound was dispersed to prepare a 10000-fold diluted powder and a
100000-fold diluted powder. These powders were added to the AIN93G
feed at 2%, to prepare a test sample 1 and a test sample 2.
[0147] That is, a test sample 1 containing Compound 1 (lophenol
compound) and Compound 2 (cyclolanostane compound) at a total
amount of 0.00002% by mass, and a test sample 2 containing these
compounds at a total amount of 0.000002% by mass were produced.
Example 1
[0148] In the present Example, using a hairless mouse which had
been ovariectomy (OVX)-treated, which is a model triggering an
imbalance of sex hormones, the osteogenesis promoting action of a
composition containing Compound 1 and Compound 2 was studied.
(1) Preparation of Sample
[0149] The test sample 1 and the test sample 2 obtained in
Production Example 3, as well as a sample containing Compound 1
(lophenol compound) alone at 0.00002% by mass (test sample 3) and a
sample containing Compound 2 (cyclolanostane compound) alone at
0.00002% by mass (test sample 4) were used. Further, a sample
containing Compound 1 (lophenol compound) alone at 0.00001% by mass
(test sample 5) and a sample containing Compound 2 (cyclolanostane
compound) alone at 0.00001% by mass (test sample 6) were also
prepared. In addition, as a control sample, an animal feed AIN93G
was used.
(2) Test Method
[0150] Female Hos: HR-1 mice which had been subjected to
ovariectomy operation (OVX) at 8 week-old (OVX mice), and female
Hos: HR-1 mice which had been subjected to sham operation (Sham) at
8 week-old (Sham mice) were purchased at 9 week-old from Japan SLC,
Inc. Both of OVX mice and Sham mice were pre-reared with an animal
feed AIN93G for 2 weeks, a control sample (AIN93G) was given to
Sham mice (6 animals) (Sham group (negative control)), OVX mice
were divided into a total of 5 groups, one group consisting of 6
animals, a control sample was given to one group (OVX group
(positive control)), a test sample 1, a test sample 2, a test
sample 3, or a test sample 4 was given to remaining four groups,
respectively (OVX+test sample group), and animals were reared for 8
weeks.
[0151] At completion of a rearing term, blood was collected from
each mouse to obtain serum, a bone-specific alkaline phosphatase
(BALP) value in blood was measured using ELISA Kit of Company
CUSABIO, and the action of the cyclolanostane compound or the
lophenol compound on osteogenesis was assessed.
(3) Test Result
[0152] The bone-specific alkaline phosphatase concentration in
blood at 8 weeks from start of administration is shown in Table 1.
In Table, a value is shown as average of one group of 6
animals.+-.standard deviation, and a p value in Table shows a
significance probability by Student t-test.
[0153] As shown in Table 1, as compared with the Sham group being a
negative control, in the OVX group being a positive control, the
bone-specific alkaline phosphatase concentration in blood showed a
low value. From this, it was seen that by ovariectomy, hormones are
deficient, the hormone balance is disturbed, and reduction in the
bone-specific alkaline phosphatase concentration in blood
occurs.
[0154] On the other hand, in the OVX group to which the test sample
1 had been given, the bone-specific alkaline phosphatase
concentration in blood showed a significantly high value as
compared with the OVX group being a positive control. Thereby, it
was seen that by ingesting a composition containing the
cyclolanostane compound and the lophenol compound in the state
where the bone-specific alkaline phosphatase concentration in blood
is reduced due to ovariectomy, the bone-specific alkaline
phosphatase concentration in blood is increased. In addition, also
in the OVX group to which the test sample 2 had been given, the
same effect as that of the OVX group to which the test sample 1 had
been given was confirmed, although not included in Table 1.
[0155] Further, also concerning the bone-specific alkaline
phosphatase concentration in blood in the OVX group to which the
test sample 3 or the test sample 4 had been given, the same degree
of the effect as that of test sample 1 or the test sample 2 was
confirmed, in every case (data not shown).
[0156] Further, also concerning the bone-specific alkaline
phosphatase concentration in blood in the OVX group to which the
test sample 5 or the test sample 6 had been given, the same degree
of the effect as that of test sample 1 or the test sample 2 was
confirmed, in every case (data not shown).
TABLE-US-00001 TABLE 1 Bone-specific alkaline phosphatase
concentration in blood Test group BALP (ng/ml) p value Sham 2967.24
.+-. 262.64 OVX 2591.07 .+-. 352.17 0.07362882 vs Sham OVX + test
sample 1 3308.01 .+-. 529.43 0.02274307 vs OVX
Example 2
[0157] In the present Example, stress load of ultraviolet
irradiation was given to a hairless mouse which had been
ovariectomy (OVX)-treated, and the anti-neuropsychiatric symptom
action of a composition containing Compound 1 and Compound 2 was
studied.
(1) Preparation of Sample
[0158] The control sample used in Example 1, the test sample 1 and
the test sample 2 produced in Production Example 3, as well as the
test sample 3 and the test sample 4 were used.
(2) Test Method
[0159] After OVX mice and Sham mice were pre-reared with an animal
feed AIN93G for 2 weeks from 9 week-old, the control sample
(AIN93G) was given to Sham mice (6 animals) (Sham group (negative
control)), OVX mice were divided into a total of five groups, one
group consisting of 6 animals, the control sample was given to one
group (OVX group (positive control)), the test sample 1, the test
sample 2, the test sample 3, or the test sample 4 was added to the
remaining four groups, respectively (OVX+test sample group), and
animals were reared for 8 weeks. Further two groups were made, one
group consisting of 6 OVX mice, the test sample 5 or the test
sample 6 was given, respectively (OVX+test sample group), and
animals were reared for 8 weeks.
[0160] For each group, after completion of pre-rearing, stress
loading (ultraviolet irradiation) was performed three times per
week, for 6 weeks from 3 weeks from start of rearing with the
control sample or the test sample. Ultraviolet irradiation was
initiated at a dose of 50 mJ/cm.sup.2, a dose was stepwisely
increased to 125 mJ/cm.sup.2, and a final total irradiation dose
was 1.7 J/cm.sup.2.
[0161] Twenty-four hours after final irradiation, blood was
collected to obtain serum, and a corticosterone value in blood was
measured using ELISA Kit of Company Enzo.
(3) Test Result
[0162] The corticosterone concentration in blood 24 hours after
final irradiation is shown in Table 2. In Table, a value is shown
as average of one group of 6 animals.+-.standard deviation and a p
value in Table shows a significance probability by Student
t-test.
[0163] As shown in Table 2, as compared with the Sham group being a
negative control, in the OVX group being a positive control, the
corticosterone concentration in blood showed a significantly high
value. From this, it was seen that by ovariectomy, hormones are
deficient, the hormone balance is disturbed, and increase in the
corticosterone concentration in blood occurs.
[0164] On the other hand, in the OVX group to which the test sample
1 or the test sample 2 had been given, the corticosterone
concentration in blood showed a significantly low value as compared
with the OVX group being a positive control. From this, it was seen
that in the state where the corticosterone concentration in blood
is increased due to ovariectomy, the corticosterone concentration
in blood can be reduced by ingesting a composition containing the
cyclolanostane compound and the lophenol compound.
[0165] Further, also concerning the corticosterone concentration in
blood in the OVX group to which the test sample 3 or the test
sample 4 had been given, in every case, the same degree of the
reducing effect as that of the test sample 1 or the test sample 2
was confirmed (data not shown).
[0166] Further, also concerning the corticosterone concentration in
blood in the OVX group to which the test sample 5 or the test
sample 6 had been given, in every case, the same degree of the
reducing effect as that of the test sample 1 or the test sample 2
was confirmed (data not shown).
TABLE-US-00002 TABLE 2 Corticosterone concentration in blood Test
group Corticosterone (ng/ml) p value Sham stress load 131.01 .+-.
39.38 OVX stress load 180.63 .+-. 43.15 0.030858 vs Sham OVX + test
sample 1 148.90 .+-. 43.00 0.199837 vs OVX stress load OVX + test
sample 2 144.77 .+-. 47.24 0.174436 vs OVX stress load
Example 3
[0167] In the present Example, the presence or absence of the
estrogen-like action of a composition containing a compound
selected from the cyclolanostane compound and the lophenol compound
as an active ingredient, in a hairless mouse which had been
ovariectomy (OVX)-treated was studied.
(1) Preparation of Sample
[0168] The control sample, the test sample 1, the test sample 2,
the test sample 3 and the test sample 4 used in Example 1 or 2 were
used.
(2) Test Method
[0169] Mice which finished rearing under the same conditions as
those of each test group of Example 2 were dissected, the uterus
was isolated at dissection, and a weight thereof was measured.
(3) Test Result
[0170] A weight of the uterine at dissection is shown in Table
3.
[0171] As compared with the Sham group being a negative control, in
the OVX group being a positive control, a significant reduction in
a weight of the uterine was recognized, but variation in a weight
of the uterine (increasing trend) was not confirmed by
administration of the test sample 1, the test sample 2, the test
sample 3, or the test sample 4, in every case. Further, this
phenomenon did not undergo influence by stress load. From the
above-mentioned result, it was suggested that the lophenol compound
and/or the cyclolanostane compound have no estrogen-like
action.
TABLE-US-00003 TABLE 3 Uterus weight Test group Uterus weight (mg)
mg/g body weight Sham 195.9 .+-. 55.7 7.18 OVX 35.2 .+-. 9.8 1.09
OVX + test sample 37.5 .+-. 12.4 1.21 Sham stress load 174.2 .+-.
55.6 6.58 OVX stress load 32.3 .+-. 11.4 0.96 OVX + test sample 1
stress load 35.8 .+-. 5.7 1.13 OVX + test sample 2 stress load 35.4
.+-. 9.0 1.12
[0172] From the above-mentioned Examples, it was seen that the
agent of the present invention is effective in improving symptoms
caused by disturbance in the sex hormone balance. Particularly, it
was seen that the agent of the present invention is effective in
improving bone dysbolism and neuropsychiatric symptom caused by
disturbance in the sex hormone balance. Further, it was suggested
that the agent of the present invention has no sex hormone-like
action. That is, it was seen that the agent of the present
invention acts on bone dysbolism and neuropsychiatric symptom, and
exerts the preventing or improving action on the symptoms. For this
reason, it was seen that the agent of the present invention is
expected to have the preventing or improving not only symptoms due
to deficiency in female hormones such as estrogen, but also the
similar symptoms due to other cause. Further, since the agent of
the present invention has no sex hormone-like action, it was also
seen that it has high safety.
Example 4
[0173] A medicament having the effect of preventing and improving
symptoms due to an imbalance of the sex hormones, consisting of the
following composition, was produced by the following method.
[0174] Two percent (2%) by mass of a composition prepared by adding
carboxymethylcellulose (CMC: manufactured by DKS Co., Ltd.) to a
mixture containing the lophenol compound produced in Production
Example 1 and the cyclolanostane compound produced in Production
Example 2 at a ratio of lophenol compound:cyclolanostane
compound=1:1 and dispersing the materials, the composition
containing 0.001% by mass of the mixture, 2% by mass of a medium
chain fatty acid (MCT: manufactured by RIKEN VITAMIN CO., LTD.), 4%
by mass of a glycerin fatty acid ester (manufactured by RIKEN
VITAMIN CO., LTD.), 0.5% by mass of saponin (manufactured by
MARUZEN PHARMACEUTICALS CO., LTD.), 0.2% by mass of ethanol
(manufactured by Japan Alcohol Corporation), 1.3% by mass of
maltitol (manufactured by HAYASHIBARA CO., LTD.), 78% by mass of
glycerin (manufactured by NOF CORPORATION) and, further, water were
added and the materials were mixed so that the total amount was
100% by mass, to produce a syrup-like preparation containing the
mixture of the lophenol compound (Compound 1) and the
cyclolanostane compound (Compound 2) at a final concentration of
0.00002% by mass.
[0175] The medicament of the present Example is effective for
improving symptoms caused by disturbance in the sex hormone
balance.
INDUSTRIAL APPLICABILITY
[0176] The present invention can be utilized for preventing or
improving symptoms caused by an imbalance of sex hormones.
* * * * *